Objective

Oral squamous cell carcinoma (OSCC) is a common malignancy with a poor prognosis. This study aimed to determine the influence and underlying mechanisms of CLSPN on OSCC.

Methods

CLSPN expression was tested using quantitative real-time polymerase chain reaction, immunohistochemistry, and western blotting. Flow cytometry, cell counting kit, and colony formation assays were performed to determine OSCC cell apoptosis, viability, and proliferation, respectively. In OSCC cells, the extracellular acidification rate (ECAR), oxygen consumption rate (OCR), glucose uptake, and lactate production were determined using the corresponding kits. Changes in the protein levels of HK2, PKM2, LDHA, Wnt3a, and β-catenin were assessed using western blotting.

Results

CLSPN expression was increased in OSCC tissues. Overexpression of CLSPN in HSC-2 cells promoted cell proliferation, increased the levels of ECAR, glucose uptake, and lactate production, and increased the protein levels of HK2, PKM2, LDHA, Wnt3a, and β-catenin, but inhibited OCR levels and apoptosis. The knockdown of CLSPN in CAL27 cells resulted in the opposite results. Moreover, the effects of CLSPN overexpression on glycolysis and OSCC cell proliferation were reversed by Wnt3a knockdown. In vivo, knockdown of CLSPN restrained tumor growth, glycolysis, and the activation of Wnt/β-catenin signaling.

Conclusion

CLSPN promoted glycolysis and OSCC cell proliferation, and reduced apoptosis, which was achieved by the activation of Wnt/β-catenin signaling pathway.

中文翻译：

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CLSPN 激活 Wnt/β-catenin 信号传导促进口腔鳞状细胞癌中的糖酵解和细胞增殖

客观的

口腔鳞状细胞癌（OSCC）是一种常见的恶性肿瘤，预后较差。本研究旨在确定 CLSPN 对 OSCC 的影响及其潜在机制。

方法

使用定量实时聚合酶链反应、免疫组织化学和蛋白质印迹测试 CLSPN 表达。进行流式细胞术、细胞计数试剂盒和集落形成测定分别测定 OSCC 细胞凋亡、活力和增殖。在 OSCC 细胞中，使用相应的试剂盒测定细胞外酸化率 (ECAR)、耗氧率 (OCR)、葡萄糖摄取和乳酸生成。使用蛋白质印迹法评估HK2、 PKM2 、LDHA、Wnt3a 和 β-连环蛋白的蛋白质水平的变化。

结果

OSCC 组织中 CLSPN 表达增加。 HSC-2细胞中CLSPN的过表达促进细胞增殖，增加ECAR水平、葡萄糖摄取和乳酸产生，并增加HK2、PKM2、LDHA、Wnt3a和β-连环蛋白的蛋白水平，但抑制OCR水平和细胞凋亡。在 CAL27 细胞中敲低 CLSPN 会产生相反的结果。此外，Wnt3a 敲低可逆转 CLSPN 过表达对糖酵解和 OSCC 细胞增殖的影响。在体内，CLSPN 的敲低抑制了肿瘤生长、糖酵解和 Wnt/β-catenin 信号传导的激活。

结论

CLSPN促进糖酵解和OSCC细胞增殖，并减少细胞凋亡，这是通过激活Wnt/β-catenin信号通路来实现的。